DK2556171T3 - Rumligt kodede biologiske assays - Google Patents

Rumligt kodede biologiske assays Download PDF

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DK2556171T3
DK2556171T3 DK11766613.1T DK11766613T DK2556171T3 DK 2556171 T3 DK2556171 T3 DK 2556171T3 DK 11766613 T DK11766613 T DK 11766613T DK 2556171 T3 DK2556171 T3 DK 2556171T3
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Mark S Chee
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Prognosys Biosciences Inc
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Claims (15)

  1. RUMLIGT KODEDE BIOLOGISKE ASSAYS
    1. Fremgangsmåde til bestemmelse af rumlige mønstre for hyppighed og/eller aktivitet for de talrige nukleinsyremål på talrige steder i en prøve, hvilken fremgangsmåde omfatter følgende trin: tilvejebringelse af en prøve, der er bundet til en bærer; tilføring af oligonukleotidsonder for talrige nukleinsyremål til de talrige steder i prøven; at lade oligonukleotidsondeme hybridisere med nukleinsyremålene; udvaskning af de ikke-hybridiserede oligonukleotidsonder fra prøven; tilføring af kodende midler på placeringer af de talrige steder i prøven ifølge et kendt rumligt mønster, hvor mindst to kodende midler tilføres til hvert af de talrige steder, og kombinationen af de kodende midler, der tilføres hvert sted, er forskellig; kobling af de kodende midler til oligonukleotidsondeme hybridiseret til nukleinsyremålene for at danne kodede sonder; bestemmelse af samtlige eller en del af en sekvens af de kodede sonder og associering af hyppighed og/eller aktivitet for de talrige biologiske mål med placeringerne af de talrige steder i prøven.
  2. 2. Fremgangsmåde til bestemmelse af rumlige mønstre for hyppighed og/eller aktivitet for de talrige biologiske mål på talrige steder i en prøve, hvilken fremgangsmåde omfatter følgende trin: tilvejebringelse af en prøve, der er bundet til en bærer; tilførsel afkodede sonder for talrige biologiske mål i et kendt rumligt mønster til de talrige steder i prøven, hvor hver kodet sonde omfatter et sondeområde, der er i stand til at interagere med et biologisk mål, og et kodende tag omfattende et oligonukleotid, der identificerer placeringen af det sted, hvor den kodede sonde tilføres; at tillade, at de kodede sonder interagerer med de biologiske mål; bestemmelse af en sekvens af de kodede sonder, hvor sekvensen omfatter det kodende tags oligonukleotidsekvens, hvorved placeringen af det sted, hvor den kodede sonde tilføres, identificeres; og associering af hyppighed og/eller aktivitet for de talrige biologiske mål med placeringerne af stederne i prøven.
  3. 3. Fremgangsmåde ifølge krav 2, hvilken fremgangsmåde endvidere omfatter efter trinnet med tilladelse og før bestemmelsestrinnet et trin med separation af de kodede sonder, der interagerer med de biologiske mål fra de kodede sonder, som ikke interagerer med de biologiske mål.
  4. 4. Fremgangsmåde ifølge krav 3, hvor de biologiske mål er nukleinsyrer, og de kodede sonder er oligonukleotidsonder.
  5. 5. Fremgangsmåde ifølge krav 1 eller 4, hvor der er to oligonukleotidsonder for hvert biologisk nukleinsyremål.
  6. 6. Fremgangsmåde ifølge krav 3, hvor de talrige biologiske mål er proteiner, sonderegioneme af de kodende sonder er proteiner og de kodende tags omfatter oligonukleotider.
  7. 7. Fremgangsmåde ifølge krav 3, hvor de talrige biologiske mål omfatter enzymer.
  8. 8. Fremgangsmåde ifølge krav 3, hvor sonderegioneme af de kodede sonder omfatter antistoffer, aptamerer, små molekyler, enzymsubstrater, formodede enzymsubstrater eller affinitetsindfangningsmidler.
  9. 9. Fremgangsmåde ifølge et hvilket som helst af kravene 1, 3, og 6, hvilken fremgangsmåde endvidere omfatter et amplifikationstrin mellem separationstrinnet eller koblingstrinnet og bestemmelsestrinnet.
  10. 10. Fremgangsmåde ifølge et hvilket som helst af kravene-1-3, hvor bestemmelsestrinnet udføres ved nukleinsyresekvensering eller high-throughput-sekvensering.
  11. 11. Fremgangsmåde ifølge krav 1, hvor koblingstrinnet udføres ved ligation eller ved forlængelse efterfulgt af ligation.
  12. 12. Fremgangsmåde ifølge et hvilket som helst af kravene 1-3, hvor produktet af de talrige biologiske mål, der testes, og de talrige steder i prøven er større end 1.000.000.
  13. 13. Fremgangsmåde ifølge et hvilket som helst af kravene 1-3 og 6, hvor sekvenserne af mindst én million kodende sonder bestemmes parallelt.
  14. 14. Fremgangsmåde ifølge et hvilket som helst af kravene 1-3, hvor det kendte rumlige mønster bestemmes ved prøvens histologiske træk.
  15. 15. Fremgangsmåde ifølge et hvilket som helst af kravene 1-3, hvor softwareprogrammeret hardware udfører mindst to trin af tilføringstrinnet, separationstrinnet, bestemmelsestrinnet og associeringstrinnet.
DK11766613.1T 2010-04-05 2011-04-05 Rumligt kodede biologiske assays DK2556171T3 (da)

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US32112410P 2010-04-05 2010-04-05
US13/080,616 US9371598B2 (en) 2010-04-05 2011-04-05 Spatially encoded biological assays
PCT/US2011/031308 WO2011127099A1 (en) 2010-04-05 2011-04-05 Spatially encoded biological assays

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US (21) US9371598B2 (da)
EP (1) EP2556171B1 (da)
JP (1) JP5893607B2 (da)
KR (1) KR101866401B1 (da)
CN (2) CN102834526B (da)
AU (1) AU2011237729B2 (da)
CA (1) CA2794522C (da)
DK (1) DK2556171T3 (da)
ES (1) ES2555106T3 (da)
HK (1) HK1222210A1 (da)
HR (1) HRP20151294T1 (da)
HU (1) HUE026666T2 (da)
PL (1) PL2556171T3 (da)
PT (1) PT2556171E (da)
RS (1) RS54482B1 (da)
SI (1) SI2556171T1 (da)
SM (1) SMT201500301B (da)
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